After blocking the membranes in TBS containing 5% BSA, 0.05% Tween 20, 0.02% NaN3 for 1 hour, primary antibodies were added in block solution for overnight incubation at 4°C.
After blocking the membranes in TBS containing 5% BSA, 0.05% Tween 20, and 0.02% NaN3 for 1 hour, primary antibodies were added in block solution for overnight incubation at 4°C.
When working characteristics were added in Block 2, age, education, monthly income, and weekly working time were significantly associated with depressive symptoms.
After adjustment for gender, age, marital status, and educational level, the independent variables ERR and over-commitment were added in block 2, before PsyCap was entered into block 3 of the model.
Working characteristics (job rank, monthly income, and weekly work time) were added in Block 2. Because job rank and monthly income are categorical variables, dummy variables were set for these two variables, respectively.
POS was added in Block 2. There were two models (Model 1 and Model 2) in Block 3. PsyCap was added in Model 1, and self efficacy, hope, resilience and optimism were added in Model 2. Asymptotic and resampling strategies were used to examine the mediating roles (a*b product) of PsyCap and its four components on the association between POS and depressive symptoms, respectively [ 35].
Alkaline phosphatase-conjugated goat anti-mouse IgG (Southern Biotechnology, Birmingham, AL) detection antibody was added in blocking buffer (1∶1,000) and incubated for 1 hr at RT.
For detection of total IgG secreting cells, 0.1 µg/well of a biotinylated polyclonal goat anti-mouse IgG (Mabtech AB) was added in blocking buffer (PBS containing 1% FCS and 0.05% Tween 20).
100 µl of antibody (diluted at a 1∶2 ratio from 5 µg/ml to 0.076 µg/ml) or clarified cell lysate (at dilutions from 1∶2 to 1∶64) was then added in blocking solution for 1 hr followed by secondary antibody, HRPO labeled polyclonal anti-human IgG Fc antibody (Roche, Switzerland) at 1∶5000 dilution, also in blocking solution, for another 1 hr.
Cells were then washed in PBS and secondary antibodies: 1∶1000 rabbit Alexa Fluor 488 and/or 1∶1000 mouse Alexa Fluor 568 (Molecular Probes) were added in blocking solution for 1 h at RT. Cells were washed again in PBS and stained with Hoechst 33342 (Molecular Probes) for 1 min. Coverslips were mounted in Prolong Gold Antifade reagent (Molecular Probes).
